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By applying this minimum read threshold, we enriched for highly expressed genes.
The minimum read threshold for each allele is one of these parameters.
Increasing the minimum read threshold required to test for an imbalance can guard against incorrect heterozygous site identification.
At the minimum read threshold of 2, a fusion was either detected or not detected in both replicates in 93% of the cases.
In addition, a minimum read threshold per allele can be applied to all heterozygous sites during imbalance detection to guard against incorrectly annotated heterozygous sites.
While it is prudent to require a minimum read threshold of reads to detect imbalances at predicted heterozygous sites, this threshold precludes the identification of complete imbalance at known heterozygous sites where only one allele is present, such as imprinted loci.
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These incorrect predictions may be partly due to sequencing errors, but as some are still present at high minimum read thresholds, errors in sequence mapping likely contribute to false positives.
Figure 4 shows the true-positive/false-positive performance curve for different minimum read thresholds compared with the other contemporary methods and our own reduced size marker library (kML) and full-sized library (kFull).
To observe the effect of changing minimum number of reads required to call a fusion, Figure 8 depicts the number of fusions detected for each replicate at different minimum reads thresholds.
To balance sensitivity and precision with incomplete genotype information, we examined the impact of changing the minimum aligned read threshold for each allele required to test for imbalanced sites.
Read-type specific raw variant lists were filtered by the SAMtools varFilter.pl utility by setting a minimum read depth threshold of six and a maximum read depth threshold of 200.
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